Developmental and Reproductive Effects of Iron Oxide Nanoparticles in Arabidopsis thaliana

Increasing use of iron oxide nanoparticles in medicine and environmental remediation has led to concerns regarding exposure of these nanoparticles to the public. However, limited studies are available to evaluate their effects on the environment, in particular on plants and food crops. Here, we investigated the effects of positive (PC) and negative (NC) charged iron oxide (Fe₂O₃) nanoparticles (IONPs) on the physiology and reproductive capacity of Arabidopsis thaliana at concentrations of 3 and 25 mg/L. The 3 mg/L treated plants did not show evident effects on seeding and root length. However, the 25 mg/L treatment resulted in reduced seedling (positive-20% and negative-3.6%) and root (positive-48% and negative-negligible) length. Interestingly, treatment with polyethylenimine (PEI; IONP-PC coating) also resulted in reduced root length (39%) but no change was observed with polyacrylic acid (PAA; IONP-NC coating) treatment alone. However, treatment with IONPs at 3 mg/L did lead to an almost 5% increase in aborted pollen, a 2%–6% reduction in pollen viability and up to an 11% reduction in seed yield depending on the number of treatments. Interestingly, the treated plants did not show any observable phenotypic changes in overall size or general plant structure, indicating that environmental nanoparticle contamination could go dangerously unnoticed.     

基于粒子群优化和支持向量机的花粉浓度预测模型

为了提高花粉浓度预报的准确率，解决现有花粉浓度预报准确率不高的问题，提出了一种基于粒子群优化（PSO）算法和支持向量机（SVM）的花粉浓度预报模型。首先，综合考虑气温、气温日较差、相对湿度、降水量、风力、日照时数等多种气象要素，选择与花粉浓度相关性较强的气象要素构成特征向量；其次，利用特征向量与花粉浓度数据建立SVM预测模型，并使用PSO算法找出最优参数；然后利用最优参数优化花粉浓度预测模型；最后，使用优化后的模型对花粉未来24 h浓度进行预测，并与未优化的SVM、多元线性回归法（MLR）、反向神经网络（BPNN）作对比。此外使用优化后的模型对某市南郊观象台和密云两个站点进行逐日花粉浓度预测。实验结果表明，相比其他预报方法，所提方法能有效提高花粉浓度未来24 h预测精度，并具有较高的泛化能力。     

Cytological Observations and Bulked-Segregant Analysis Coupled Global Genome Sequencing Reveal Two Genes Associated with Pollen Fertility in Tetraploid Rice

Neo-tetraploid rice with high fertility is a useful germplasm for polyploid rice breeding, which was developed from the crossing of different autotetraploid rice lines. However, little information is available on the molecular mechanism underlying the fertility of neo-tetraploid rice. Here, two contrasting populations of tetraploid rice, including one with high fertility (hereafter referred to as JG) and another with low fertility (hereafter referred to as JD), were generated by crossing Huaduo 3 (H3), a high fertility neo-tetraploid rice that was developed by crossing Jackson-4x with 96025-4x, and Huajingxian74-4x (T452), a low fertility autotetraploid rice parent. Cytological, global genome sequencing-based bulked-segregant (BSA-seq) and CRISPR/Cas9 technology were employed to study the genes associated with pollen fertility in neo-tetraploid rice. The embryo sacs of JG and JD lines were normal; however, pollen fertility was low in JD, which led to scarce fertilization and low seed setting. Cytological observations displayed low pollen fertility (25.1%) and approximately 31.3 and 27.2% chromosome lagging at metaphase I and II, and 28.8 and 24.8% chromosome straggling at anaphase I and II in JD, respectively. BSA-seq of F_2–3 generations and RNA-seq of F₄ generation detected a common fragment, i.e., 18,915,234–19,500,000, at chromosome 7, which was comprised of 78 genes associated with fertility. Among 78 genes, 9 genes had been known to be involved in meiosis and pollen development. Two mutants ny1 (LOC_Os07g32406) and ny2 (LOC_Os07g32040) were generated by CRISPR/Cas9 knockout in neo-tetraploid rice, and which exhibited low pollen fertility and abnormal chromosome behavior. Our study revealed that two unknown genes, LOC_Os07g32406 (NY1) and LOC_Os07g32040 (NY2) play an important role in pollen development of neo-tetraploid rice and provides a new perspective about the genetic mechanisms of fertility in polyploid rice.     

Analysis of saturated free fatty acids from pollen by HPLC with fluorescence detection

A sensitive method for the determination of free fatty acids using 2‐(2‐(anthracen‐10‐yl)‐1H‐naphtho[2,3‐d]imidazol‐1‐yl) ethyl‐p‐toluenesulfonate (ANITS) as tagging reagent with fluorescence detection has been developed. ANITS could easily and quickly label fatty acids in the presence of the K₂CO₃ catalyst at 90 °C for 40 min in N,N‐dimethylformamide solvent. From the extracts of rape bee pollen samples, 20 free fatty acids were sensitively determined. Fatty acid derivatives were separated on a reversed‐phase Eclipse XDB‐C₈ column by HPLC in conjunction with gradient elution. The corresponding derivatives were identified by post‐column APCI/MS in positive‐ion detection mode. ANITS‐fatty acid derivatives gave an intense molecular ion peak at m/z [M+H]⁺; with MS/MS analysis, the collision‐induced dissociation spectra of m/z [M+H]⁺ produced the specific fragment ions at m/z [M–345]⁺ and m/z 345.0 (here, m/z 345 is the core structural moiety of the ANITS molecule). The fluorescence excitation and emission wavelengths of the derivatives were λ_ex = 250 nm and λ_em = 512 nm, respectively. Linear correlation coefficients for all fatty acid derivatives are >0.9999. Detection limits, at a signal‐to‐noise ratio of 3 : 1, are 24.76–98.79 fmol for the labeled fatty acids.     

参芪花粉片生产工艺改进探究

为了减少工艺流程、提高生产效率和产品质量,对参芪花粉片生产工艺的发酵法破壁和灭菌两个环节进行了研究与改进.采用75%的乙醇水溶液作为花粉破壁、制粒的润湿剂,同时又兼作灭菌剂,提出了采用有机溶剂加胶体磨的新工艺方法替代直接发酵法对花粉进行破壁.通过改进工艺,主要工艺由5步减少到4步,工时显著减少,灭菌效率显著提高,生产成本降低,产品的外观和品质得到了改善;并且新工艺可提高片剂的稳定性和有效成分的均匀度.     

温差-超声波复合破壁法对花粉破壁率和黄酮得率的影响

采用温差破壁结合超声波辅助的复合花粉破壁法对花粉进行破壁,考察了不同的温差破壁因素对花粉破壁率和黄酮得率的影响,并以黄酮得率为指标进行正交试验L9（3^3）优化因素,得到温差破壁的最佳工艺条件为：温差100℃、料液比1：15、冷冻时间24h,此条件下花粉破壁率为61．61％,黄酮得率为3．166％．进一步用超声辅助对花粉进行破壁,使其破壁率达到85％～90％,黄酮得率提高到3．310％．同时,通过试验分析阐述了破壁率和黄酮得率两者的联系．     

油菜和荷花蜂花粉提取物的抗氧化性研究

采用超声法,分别以甲醇、水为溶剂提取油菜和荷花的蜂花粉活性成分,通过测定油菜和荷花蜂花粉的提取物对超氧阴离子自由基、羟基自由基的清除率及对脂质过氧化的抑制率,评价两种蜂花粉的抗氧化能力。结果表明:两种蜂花粉提取物均具有较强的抗氧化作用,油菜蜂花粉提取物比荷花蜂花粉提取物有更强的抗氧化性。油菜蜂花粉甲醇提取物对超氧阴离子自由基、羟基阴离子自由基清除率可达到83.2%,99.2%,对脂质过氧化的抑制率可达到92.2%。将蜂花粉抗氧化提取物分为甲醇部、水部溶剂萃取物,经抗氧化活性比较,其强弱顺序为甲醇部>水部。     

Possibility of Japanese Cedar Pollen Causing False Positives in the Deep Mycosis Test

Because Japanese cedar pollen (JCP) contains beta-1,3-d-glucan (BG), there is concern that its lingering presence in the atmosphere, especially during its scattering period, may cause false positives in the factor-G-based Limulus amebocyte lysate (LAL) assay used to test for deep mycosis (i.e., G-test). Hence, we examined whether the LAL assay would react positively with substances contained in JCP by using the G-test to measure JCP particles and extracts. BG was purified from the JCP extract on a BG-specific affinity column, and the percentage extractability was measured using three different BG-specific quantitative methods. The G-test detected 0.4 pg BG in a single JCP particle and 10 fg from a single particle in the extract. The percentage extractability of JCP-derived BG was not significantly different among the three quantitative methods. As the JCP particles should technically have been removed during serum separation, they should be less likely to be a direct false-positive factor. However, given that the LAL-assay-positive substances in the JCP extract were not distinguishable by the three BG-specific quantitative methods, we conclude that they may cause the background to rise. Therefore, in Japan false positives arising from JCP contamination should be considered when testing patients for deep mycosis.